TY - JOUR
T1 - SETD1A methyltransferase is physically and functionally linked to the DNA damage repair protein RAD18
AU - Alsulami, Manal
AU - Munawar, Nayla
AU - DIllon, Eugene
AU - Oliviero, Giorgio
AU - Wynne, Kieran
AU - Alsolami, Mona
AU - Moss, Catherine
AU - Gaora, Peadar
AU - O'Meara, Fergal
AU - Cotter, David
AU - Cagney, Gerard
N1 - Publisher Copyright:
© 2019 Alsulami et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2019
Y1 - 2019
N2 - SETD1A is a SET domain-containing methyltransferase involved in epigenetic regulation of transcription. It is the main catalytic component of a multiprotein complex that methylates lysine 4 of histone H3, a histone mark associated with gene activation. In humans, six related protein complexes with partly nonredundant cellular functions share several protein subunits but are distinguished by unique catalytic SET-domain proteins. We surveyed physical interactions of the SETD1A-complex using endogenous immunoprecipitation followed by label-free quantitative proteomics on three subunits: SETD1A, RBBP5, and ASH2L. Surprisingly, SETD1A, but not RBBP5 or ASH2L, was found to interact with the DNA damage repair protein RAD18. Reciprocal RAD18 immunoprecipitation experiments confirmed the interaction with SETD1A, whereas size exclusion and protein network analysis suggested an interaction independent of the main SETD1A complex. We found evidence of SETD1A and RAD18 influence on mutual gene expression levels. Further, knockdown of the genes individually showed a DNA damage repair phenotype, whereas simultaneous knockdown resulted in an epistatic effect. This adds to a growing body of work linking epigenetic enzymes to processes involved in genome stability.
AB - SETD1A is a SET domain-containing methyltransferase involved in epigenetic regulation of transcription. It is the main catalytic component of a multiprotein complex that methylates lysine 4 of histone H3, a histone mark associated with gene activation. In humans, six related protein complexes with partly nonredundant cellular functions share several protein subunits but are distinguished by unique catalytic SET-domain proteins. We surveyed physical interactions of the SETD1A-complex using endogenous immunoprecipitation followed by label-free quantitative proteomics on three subunits: SETD1A, RBBP5, and ASH2L. Surprisingly, SETD1A, but not RBBP5 or ASH2L, was found to interact with the DNA damage repair protein RAD18. Reciprocal RAD18 immunoprecipitation experiments confirmed the interaction with SETD1A, whereas size exclusion and protein network analysis suggested an interaction independent of the main SETD1A complex. We found evidence of SETD1A and RAD18 influence on mutual gene expression levels. Further, knockdown of the genes individually showed a DNA damage repair phenotype, whereas simultaneous knockdown resulted in an epistatic effect. This adds to a growing body of work linking epigenetic enzymes to processes involved in genome stability.
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U2 - 10.1074/mcp.RA119.001518
DO - 10.1074/mcp.RA119.001518
M3 - Article
C2 - 31076518
AN - SCOPUS:85068798698
SN - 1535-9476
VL - 18
SP - 1428
EP - 1436
JO - Molecular and Cellular Proteomics
JF - Molecular and Cellular Proteomics
IS - 7
ER -